狐狸源肺炎克雷伯菌的分离鉴定及毒力检测  

Isolation,identification and virulence factors detection of Klebsiella pneumoniae isolated from fox

作　　者：韩坤[1] 孟祥玉[1] 白雪[1] 闫喜军[1] 冯二凯[1] 佟盼盼 吕娇 于莹 陈立志[1,4] HAN Kun;MENG Xiang-yu;BAI Xue;YAN Xi-jun;FENG Er-kai;TONG Pan-pan;LYU Jiao;YU Ying;CHEN Li-zhi(Institute of Special Animal and Plant Science of CA.AS,Changchun 130122,China;College o f Veterinary Medicine、Xi?ij icing Agricultural University,Wulumuqi 130118, China;College of Animal Science and Techiiology,Jilin Agricultural University,Changchun 130118, China;Jilin Special Research Biological Technology Co,Ltd , Changchun 130122,China)

机构地区：[1]中国农业科学院特产研究所，吉林长春130122 [2]新疆农业大学动物医学学院，新疆乌鲁木齐830052 [3]吉林农业大学动物科学技术学院,吉林长春130118 [4]吉林特研生物技术有限责任公司，吉林长春130122

出　　处：《中国兽医学报》2019年第4期722-727,共6页Chinese Journal of Veterinary Science

基　　金：吉林省重点科技公关计划资助项目(20150204073NY)

摘　　要：从濒死期蓝狐肺脏中分离到1株革兰阴性短杆菌,并对分离菌株进行形态学观察、16S rRNA鉴定、全自动细菌生化鉴定仪鉴定及khe基因鉴定,在此基础上,完成了分离菌株的分型、人工感染小鼠试验、药敏试验及毒力基因检测等。最终鉴定结果显示,该分离菌株为K20型肺炎克雷伯菌,命名为SWKp17;同时,Vitek细菌鉴定仪鉴定该菌为肺炎克雷伯菌。人工感染试验和药敏试验结果表明,该细菌有致病性和多重耐药性。其对小鼠的半数致死量(LD_(50))为2.9×10~7 CFU;对三代头孢菌素类、氯霉素类、喹诺酮类、氨基糖苷类、四环素类、磺胺类耐药,但对亚胺培南和多黏菌素敏感。该菌株具有fim、ureA、mrkD及wabG毒力基因,但不具有rmpA、kfu、alls、iroNB、ybtA、uge、iucB等毒力基因。结果表明,引起本次狐狸肺炎的致病菌为非K1、K2、K3、K5、K20、K54及K57型肺炎克雷伯菌,本研究为临床治疗由该菌引起的狐狸肺炎提供参考依据。In order to identify the pathogen caused a serious mortality of blue fox in Shandong Province,the lung samples from the dying blue fox were collected and one gram-negative bacterium was isolated and purified.Then carried out the 16 S rRNA identification,automatic biochemical identification of bacteria identification and khe gene identification.On this basis,the serotype characteristics,pathogenicity tests,drug sensitivity test as well as genetic evolution analysis of main virulence genes were completed.16 S rRNA and khe gene sequence identification results showed that the isolated strain K54 was Klebsiella pneumoniae,named as SWKp17;the same time,Vitek bacterial identification instrument identified as Klebsiella pneumoniae.Pathogenicity tests and drug sensitivity test results showed that the strain was pathogenic and multi-drug resistance.The median lethal dose(LD50) in mice was 2.9×107 CFU.The antimicrobial sensitivity test showed that the pathogen was resistant to third-generation cephalosporins,chloramphenicol,quinolones,aminoglycosides,tetracyclines and sulfonamides,sensitive to imipenem and polymyxin.It contained fim,ureA,mrkD and wabG genes,but did not contain virulence rmpA,kfu,alls,iroNB,ybtA,uge and iucB genes.The results of this study confirmed that the died fox did not caused by K1,K2,K3,K5,K20,K54 and K57 Klebsiella pneumoniae.These results provided a reference for clinical treatment of fox pneumonia caused by the bacteria.

关 键 词：狐狸 肺炎克雷伯菌 分离 鉴定 毒力基因 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]